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Internalized ubiquitinated UT-A1 accumulated mainly in the early endosome (EE). UT-A1-MDCK cell were treated without or with 10 μM FSK for 3 h alone or together with chlorpromazine (Chlorp, 10 μg/ml, pretreated for 1 h), then the cells were subjected to EE isolation. The lysates from the whole cells and EE were blotted with UT-A1 antibody (A) or immunoprecipitated by UT-A1 antibody, then detected by ubiquitin antibody (B). These results were reproduced two additional times.
